Necrotic Lesions Research Articles

Diagnostic Challenges of Mucormycosis in a Patient with Diabetes: A Case Report

Mucormycosis is a rare and life-threatening fungal infection that predominantly affects immunocompromised individuals, particularly those with poorly controlled diabetes mellitus. Achieving an accurate diagnosis remains a significant challenge in clinical practice. We report a case of mucormycosis in a 25-year-old woman who presented with an altered mental status and a 5 × 2 cm necrotic lesion on the left nasal septum, characterized by indistinct borders, during admission. Prolonged diagnostic investigations ultimately led to the identification of nasal mucormycosis attributable to Rhizomucor pusillus, a rare species within the Mucorales order, which complicates the diagnostic process. Unfortunately, the patient's clinical condition rapidly deteriorated, resulting in her demise prior to the administration of systemic antifungal therapy. Although the comprehensive evaluations facilitated the identification of the causative agent, the time required for this process did not favor the patient's prognosis. The atypical symptoms, swift progression, and infrequent occurrence of mucormycosis underscore the critical importance of its timely and accurate diagnosis to ensure favorable patient outcomes. Proper identification of the pathogen and immediate administration of therapeutic interventions can considerably enhance the likelihood of patient recovery and survival.

Bronchial washing fluid sequencing is useful in the diagnosis of lung cancer with necrotic tumor

BackgroundEarly-stage lung cancers detected by low-dose computed tomography (CT) often require confirmation through invasive procedures due to the absence of endobronchial lesions. This study assesses the diagnostic utility of bronchial washing fluid (BW) sequencing, a less invasive alternative, aiming to identify patient characteristics most suited for this approach. MethodsFrom June 2017 to March 2018, we conducted a prospective cohort study by enrolling patients with incidental lung lesions suspected of early-stage lung cancer at two independent hospitals, and 114 were diagnosed with lung cancer while 50 were diagnosed with benign lesions. BW sequencing was performed using a targeted gene panel, and the clinical characteristics of patients detected with cancer through sequencing were identified. ResultsMalignant cells were detected in 33 patients (28.9 %) through BW cytology. By applying specificity-focused mutation criteria, BW sequencing classified 42 patients (36.8 %) as having cancer. Among the cancer patients who were BW sequencing positive and BW cytology negative, 15 patients (75.0 %) showed necrosis on CT. The sensitivity of BW sequencing was particularly enhanced in patients with necrotic tumors, reaching 75 %. ConclusionsBW sequencing presents a viable, non-invasive diagnostic option for early-stage lung cancer, especially valuable in patients with necrotic lesions. By potentially reducing the reliance on more invasive diagnostic procedures, this method could streamline clinical workflows, decrease patient burden, and improve overall diagnostic efficiency.

First Report of Root Collar Canker Caused by Neoscytalidium dimidiatum on Jacaranda mimosifolia in China.

Jacaranda mimosifolia is widely cultivated as a garden ornamental tree. In July 2023, an unknown root collar canker of J. mimosifolia was discovered in green belts of Qingxiu District, Nanning, China, with a 8% incidence rate. Crowns of affected trees ranged from reddish brown leaves to deciduous or dead. Root collar tissue became necrotic matched by underbark dark brown lesions with irregular margins, and rotted at last. Six diseased plants distributed within 3000 m2 were choosed, and 24 root collar tissues were surface sterilized and placed on potato dextrose agar (PDA) plates to incubate at 28℃ for 3 to 5 days. Same colonies were consistently isolated from 18 tissues, and three isolates (M3-B1-1, M3-B1-2 and M3-B1-3) were purified for morphological and molecular determination. These isolates formed colonies with lush aerial mycelia rapidly, which covered a 90 mm plate in 72h. The colonies were initially white, then grayish-green to black. Arthrospores were colourless to light brown, short columnar, aseptate, truncate base, averaging 12.1±2.5 µm × 3.4±0.7 µm, sometimes formed arthric chains. Chlamydospores were dark brown, round or oval, aseptate, averaging 8.7±1.6 µm × 5.0±0.9 µm. Mature pycnidia and conidia produced for about 50 days on oatmeal agar medium (OMA), and conidia were colorless, oblong, aseptate, averaging 11.2±1.2 µm × 6.0±1.4 µm. These morphological characteristics were consistent with the description of Neoscytalidium dimidiatum (Penz.) Crous & Slippers (Crous et al. 2006). Genomic DNA was extracted from three isolates. The partial ITS region, TUB2 and TEF1-α genes were amplified (White et al., 1990; Glass and Donaldson 1995; Carbone and Kohn 1999). The sequences were deposited in GenBank (ITS: PP939650-PP939652; TUB2: PP942728-PP942730; TEF1-α: PP942731-PP942733). Blastn analysis revealed that ITS sequences of three isolates showed 99.8%, 100%, 100% identity (506 bp out of 507 bp, 507 bp out of 507 bp, 507 bp out of 507 bp) to N. dimidiatum C21 (KX447539), the TUB2 sequences showed 100% identity (436 bp out of 436 bp, 437 bp out of 437 bp, 437 bp out of 437 bp) to N.dimidiatum LNeo (ON099066), and the TEF1-α sequences showed 99.64% identity (276 bp out of 277 bp) to N.dimidiatum ARM230 (MK495384), respectively. Phylogenetic analysis based on concatenated ITS, TUB2 and TEF1-α sequences showed that three isolates were clustered into the same clade as N. dimidiatum. To fulfill Koch's postulates, pathogenicity of these isolates was tested on healthy two-year-old J. mimosifolia trees. Stem and root collar were wounded and placed mycelial plugs (8mm), and the inoculation sites were wrapped with parafilm or covered with nursery substrate to maintain the humidity. Four plants were inoculated with each isolate. As a control, four plants were inoculated with noncolonized PDA plugs. All treated plants were kept in a greenhouse at 28 ± 3°C and 70% relative humidity. Foliar blight and necrotic lesions around inoculation points were observed about 65 days after inoculation, and 50% of inoculated trees exhibited symptoms, whereas the control trees remained symptomless. Neoscytalidium dimidiatum was successfully reisolated from symptomatic tissue via morphological analysis. To our knowledge, this is the first report of root collar canker caused by N. dimidiatum on J. mimosifolia. Neoscytalidium dimidiatum has a wide range of hosts, including pitaya, pine, mulberry, pear, grape, locust tree (Luo et al. 2024). This finding will help in controlling of the disease epidemic.

Genetic identification of methicillin resistant Staphylococcus aureus (MRSA) isolated from diabetic foot ulcers and evaluating the inhibition activity of reuterin against this bacteria.

To genetically diagnose the isolates of methicillin-resistant staphylococcus aureus taken from patients with severe diabetic foot infections, and to test the inhibitory effect of reuterin on the growth of these bacteria. The experimental study was conducted from June to November 2021 at the Diabetes Unit of Al-Faihaa General Hospital, Basrah, Iraq, and comprised 62 foot ulcer swabs from the necrotic lesions of patients with type 2 diabetes. The swabs were cultivated first in brain heart infusion broth media, and then streaked on Mannitol salt agar and staphylococcus chromogeneic agar media for phenotypic and genetic analysis. The genetic identification of bacteria was confirmed by deoxyribonucleic acid extraction, and methicillin-resistant staphylococcus aureus was confirmed by the presence of plasmid mecA gene. The inhibition activity of reuterin towards methicillin-resistant staphylococcus aureus was determined using the minimum inhibitory concentration test. All data was analyzed with SPSS version 23. A total of 62 swabs were taken from the necrotic lesions of type 2 diabetes mellitus (T2DM) patients with diabetic foot ulcers. Of the total isolates, 9(14.5%) gave mauve to purple colour on staphylococcus chromogeneic agar, which was then genetically confirmed as methicillin-resistant staphylococcus aureus. The minimum inhibitory concentration value of these bacteria was 10μl/ml at 16mm inhibition zone diameter. There was no cytotoxicity of reuterin to human red blood cells. Reuterin was found to be a natural antimicrobial substance suitable for use to disinfect diabetic foot wounds from bacterial contamination and infection, especially those caused by methicillin-resistant staphylococcus aureus.

Thromboangiitis Obliterans (Buerger's Disease): A Young Woman's Rare Case and Current Knowledge

AbstractIn this paper, we described the clinical features and the typical vascular imaging picture of a rare case of thromboangiitis obliterans (TAO) in a 47-year-old, young, Caucasian woman without relevant medical records except for smoking habits and admitted to our Department of Internal Medicine with a necrotic lesion of the first and third toes of the right foot. After a rapid clinical and instrumental evaluation, necrotic tissue debridement was performed and the patient was treated with Iloprost infusion and antibiotics (for overlapping infection) with clinical improvement. Moreover, we summarized the current knowledge about the disease making an update and critical examination of the literature. TAO, also known as Buerger's disease, is a rare disease. In recent decades, the incidence of TAO has decreased in Europe and the United States in men but several studies have reported an increase in the prevalence of disease in women. TAO is a segmental nonatherosclerotic inflammatory disorder that involves primarily small and medium arteries, veins, and nerves of the extremities with limb ischemia, pain, claudication, ulcers, and necrosis that can lead to amputation. Smoking is considered the main risk factor, but despite that, more than 100 years have passed since the first description of the disease by Leo Buerger, and knowledge on pathogenesis, precipitating factors, and therapy remain still unclear.

Chlorine Dioxide: Antiviral That Reduces the Spread of ToBRFV in Tomato (Solanum lycopersicum L.) Plants.

Tomato brown rugose fruit virus (ToBRFV), being a mechanically transmitted disease, is usually difficult to control; therefore, an effective alternative to reduce transmission and replication in the crop is by spraying with chlorine dioxide (ClO2) during routine crop management. In this research, the efficacy of chlorine dioxide (ClO2) for ToBRFV management in a greenhouse and open field was determined. The phytotoxicity of ClO2 and its effective concentration against ToBRFV in Nicotiana longiflora plants were evaluated. Subsequently, the effect of ClO2 on ToBRFV was evaluated in tomato plants grown in an open field. Finally, the effectiveness of ClO2 on plants inoculated with ToBRFV under greenhouse conditions was evaluated and the number of necrotic local lesions (NLLs) was quantified. The results revealed that ClO2 at 760 mg L-1 did not show phytotoxicity and reduced the number of NLLs in N. longiflora plants. It also decreased ToBRFV transmission and replication in field- and greenhouse-grown tomato plants, improving agronomic parameters. ClO2 reduced replication in plants inoculated with different amounts of ToBRFV inoculum in a greenhouse. N. longiflora leaves expressed lower numbers of NLLs when inoculated with ClO2-treated tomato plant extracts. Finally, the results demonstrate that ClO2 represents an effective management alternative when used by direct application to plants. To our knowledge, this is the first study where the use of an antiviral compound is carried out under field and greenhouse conditions.

Trash Feet and Mesenteric Ischemia as a Rare Manifestation of a Complicated Abdominal Aortic Aneurysm: A Case Report.

Abdominal aortic aneurysms and atheromatous plaques are well-established potential culprits in the development of cholesterol embolization. This embolic phenomenon can result in the occlusion of peripheral arteries in the forefoot or toes, commonly referred to as "trash foot". A 61-year old patient exhibited severe symptoms and signs of lower limb and mesenteric ischemia, represented clinically by abdominal pain, hematochezia, and bilateral plantar necrotic lesions, attributed to embolization from a previously unknown large infrarenal abdominal aortic aneurysm. Diagnosis was confirmed by performing angio-CT of the abdomen, an angiologic examination, as well as colonoscopy. The patient underwent open surgical repair of the aortic aneurysm and received intravenous prostanoids, resulting in a successful outcome with resolution of bowel ischemia and complete recovery of bilateral foot lesions with no tissue loss. Cholesterol embolization syndrome seldom manifests with its complete clinical spectrum. Identifying and addressing the underlying cause is crucial, with a focus on potential abdominal aortic aneurysms. A combined approach involving surgical repair and prostanoid therapy may offer a viable option for patients experiencing severe manifestations.

Clinical and morphological specifics, pathogenetic ways of skin manifestations in SARS-CoV-2 infection

To date, the most studied clinical and morphological changes characteristic of SARS-CoV-2 virus are its pulmonary manifestations and cardiovascular involvement. However, the evaluation of COVID-associated skin changes and the analysis of their mechanisms are also important for us, because the skin manifestations of the infection can change the patient's appearance for the worse, i.e. affect the aesthetic sphere and significantly reduce the quality of life. The study of not only skin manifestations of COVID-19, but also their morphological substrate and pathogenetic basis allows us to apply the most effective methods of treatment and provide adequate management of patients even in the post-coital status. According to the information collected to date, the most frequently reported skin manifestations of SARS-CoV-2 are pseudo-frostbite, maculopapular and vesicular lesions, urticaria, lividoid and necrotic lesions, hemorrhagic purpura (vasculitis), and conditions from the group of other unclassified skin lesions. Despite the variety of clinical variants of SARS-CoV-2-associated skin changes, morphological stigmas are often stencil-like: these are areas of lymphohistiocytic infiltration of perivascular localization, the presence of fibrinoid necrosis foci in the vessel walls, the formation of occlusive thrombi, erythrocyte extravasation. Mechanisms of damage to epidermis and dermis within COVID-19 infection may be due to the influence of complement components, activation of cytotoxic lymphocytes and NK-cells, excessive synthesis of proinflammatory cytokines, in particular, interleukin 6, as well as interferons, hyperergic reactions. In addition to routine morphological, immunohistochemical examination of skin biopsy specimens from patients with different forms of skin manifestations of SARS-CoV-2 infection is an important tool in the diagnostic confirmation of COVID-associated dermatologic pathology, especially in patients with a suspected history of this disease or with problematic laboratory results.

Three-dimensional mapping of necrotic lesions for early-stage osteonecrosis of the femoral head

BackgroundThere is a scarcity of evidence regarding the potential relationship between the size and location of necrotic lesions, which must be understood to provide optimal joint-preserving treatment. The purpose of this study was to characterize the distribution patterns of necrotic lesions of varying sizes in early-stage osteonecrosis of femoral head (ONFH) with the use of three-dimensional mapping.MethodsWe retrospectively evaluated clinical CT images of the hips that were performed in the Third Hospital of Hebei Medical University from January 2018 to December 2022 and collected all CT images diagnosed with stage I and II ONFH. Three-dimensional structures that included both necrotic lesions and normal areas of the femoral heads were reconstructed and divided into eight regions to record their size and location. CT images for all lesions were superimposed onto a standard template, and three-dimensional mapping was created to determine the presence of concentrated areas of lesions.ResultsIn a cohort of 143 patients with stage I and II ONFH, a total of 150 hips were reviewed. For lesions with less than 15% of the femoral head volume, necrotic lesions predominantly involve regions I, III, and V, with region I showing concentration. For lesions with volumes ranging from 15 to 30%, necrotic lesions exhibited a wider distribution across regions I, II, III, IV, V, and VII, with significant concentrations in regions I, III, and V. For lesions exceeding 30% of the femoral head volume, the necrotic lesions were extensively distributed across nearly the entire femoral head, with a notable expansion of the concentrated necrotic areas.ConclusionsThe distribution of necrotic lesions varies with lesion size, with smaller lesions primarily concentrated in the anterior and medial regions of the femoral head, particularly in the anterosuperior region, while larger lesions expand to the lateral and inferior regions. These findings enhance existing classification systems and provide crucial insights for guiding hip-preserving surgical planning and approaches.

First Report of Fusarium avenaceum Causing Root Rot of Raspberry (Rubus corchorifolius) in China.

Raspberry (Rubus corchorifolius) plants hold historical, economic, and medicinal importance in China (Yang et al. 2022). Raspberries are cultivated to generate income for local farmers in Lintao County, Dingxi City, Gansu Province. However, farmers encountered challenges due to raspberry plants exhibiting root rot disease, resulting in plant death. During a thorough field survey conducted in June 2022, symptoms ranging from leaf yellowing and wilting to necrotic lesions and root rots were observed, where approximately 30% of raspberry plants were affected. Five diseased and healthy plants were collected from the farmers' fields in Lintao (35.53oN, 103.84oE) for pathogen identification. Symptomatic and asymptomatic root tissues were surface sterilized with 75% ethanol for 30 s and 3% NaOCl for 5 min, followed by three rinses in sterile water. Small pieces (0.5 × 0.5 cm) were cut and incubated on potato dextrose agar (PDA) plates at 25°C for 7-10 days. Twenty-two pure Fusarium isolates, which displayed four distinct colony groups morphologically, were obtained. Pathogenicity tests on isolates RB10, RB1, RB30, and RB23, representing each colony group, revealed that RB10 exhibited symptoms similar to those observed in the field. The RB10 strain produced yellowish-white to greyish-white colonies on PDA and was then cultured in a carboxymethylcellulose (CMC) broth for enhanced conidia production (Zhang et al. 2020). Macroconidia were sickle-shaped or slightly curved, with three to five septa (19.2 to 38.5 x 3.1 to 5.8 µm, n =40). Microconidia were oval to ellipsoidal, non-septate or featuring 1 to 2 septa (4.8 to 10.5 x 2.1 to 5.2 µm, n=20). These morphological features indicated the isolate was similar to Fusarium avenaceum (Leslie and Summerell, 2006). For further identification of the strains, genomic regions (ITS-rDNA, TEF-1α, and RPB2) were amplified and sequenced using specific primers ITS1/ITS4, EF-1/EF-2, and 5f2/7cr, respectively (O'Donnell et al. 2010; Uwaremwe et al. 2021; Zarrin et al. 2016). PCR BLASTn queries of NCBI GenBank revealed a 99.8% (522 bp), 99.4% (355 bp) and 99.6% (985 bp) homology with F. avenaceum (MZ724839.1, MN271631.1, and MK185026.1), respectively. Sequences were deposited in GenBank (ITS, OR735571; TEF-1α, PP216660; RPB2, PP857820). One-year-old raspberry seedlings were planted in pots with a sterile soil mix (2:2:1 v/v ratio of soil, peat, and vermiculite) under controlled greenhouse conditions (23-26°C, 16h light/8h dark). A month post-planting, taproots were wounded in six pots and inoculated with 20 ml of conidia suspension (106 conidia/ml), while the other six pots were maintained as controls. After 14 days, RB10-infected plants showed symptoms similar to field observations, while controls remained healthy. The experiment was conducted twice, and re-isolation confirmed both the pathogenicity and identity of the pathogen. In the concatenated phylogenetic tree of ITS, TEF-1α and RPB2, strain RB10 was clustered with the F. avenaceum representative strains KG502, KG431 and F094. Studies revealed F. avenaceum varied pathogenicity across plants (Bugingo, 2022; Moparthi et al. 2020& 2024; Yli-Mattila et al. 2018), and it has been reported to induce raspberry fruit rot (Wang et al. 2017). However, no previous reports linked this fungus to raspberry root rot. This report is crucial for understanding the impact of root rot disease on raspberry cultivation and developing effective management strategies.

First Report of Stem Blight of Mungbean (Vigna radiata) Caused by Diaporthe longicolla in the United States.

Mungbean (Vigna radiata) is primarily grown in Asia and directly consumed by humans. U.S. consumers embraced mungbean as a plant-based protein in vegan eggs and meat substitutes. New cultivars are being developed for American farmers because of the crop's tolerance to heat and drought, and its adaptability to current farming infrastructure. Mungbean's short season complements various cropping systems such as intercropping, alternative cropping, and green manure. With rotations and inclusion with soybean systems, there is a concern about the overlap of common pathogens for soybean and mungbean. During August 2022 when mungbeans reached full maturity (growth stage R6), reddish-brown and necrotic stem lesions with linear rows of black pycnidia were observed on Berken and OK2000 cultivars at fields located in Hancock County, IA and Story County, IA in the United States. Pycnidia measured 0.5-0.6mm in length. Disease incidence was approximately 10% of plants in Hancock County, IA and less than 3% of plants in Story County, IA. Pycnidia from 16 plants were excised and immersed in a 0.5% NaOCl solution for 1 min, rinsed with autoclaved distilled water, and placed onto potato dextrose agar (PDA). Eighteen isolates were hyphal tipped and grown on PDA and were stored at 25°C. Isolates were then visually identified by culture and conidia morphology (Hobbs et al. 1985, Santos et al. 2011). Colonies were cream to white, dense, and floccose. Large black stromata were formed in a concentric pattern or scattered; alpha conidia were ellipsoidal. Template DNA for PCR amplification of the internal transcribed spacer region of the nuclear ribosomal DNA operon (ITS) and the beta-tubulin gene (TUB) was extracted from 18 isolates by scraping mycelia with a sterile pipette tip and transferring it into 50 ul of PrepMan Ultra Sample Reagent (Applied Biosystems, Foster City, California, USA). Fungal primers were ITS1 and ITS4 (White et al. 1990) and Bt-2F/Bt-2R (Udayanga et al. 2014). Sequences of isolates obtained from fields in both counties were identical, providing no species diversity. GenBank accession numbers for the ITS region were PP105598 and PP105599; PP108254 and PP108255 for TUB sequences. BLAST results showed the ITS 550/550 base pairs with type specimen D. longicolla ATCC 60326 GB NR_144924 and the TUB 446/446 base pairs with type specimen D. longicolla ATCC 60325 GB KJ610883. Thus, the isolates were identified as Diaporthe longicolla (Hobbs) J.M. Santos, Vrandecic & A.J.L. Phillips based on morphology and molecular characters (Santo et al 2011; Udayanga et al. 2014). To confirm the pathogenicity of the D. longicolla isolates, twenty mungbean plants (cv. Berken and OK2000) were grown in the greenhouse at 85% RH and 16hr light for 20 days. Inoculum was prepared by placing sterile toothpicks on 1/3 PDA with a single representative isolate from each field location for 21 days (Ghimire et al. 2019). Mungbean plants were grown in a 10cm-by-10cm pot containing a greenhouse professional growing mix (Sungrow, Agawam, Massachusetts, USA) and grown for 30 days post emergence. After 12 days of growing, a 3mm segment of the infested toothpick was inserted into a stem wound below the first trifoliate and sealed with parafilm. A sterile toothpick was inserted into the control plants. After 14 days, red lesions extended downward 1 to 3 cm from the inoculation site, and white mycelial was present in the wound. At 21 days red lesions spanned 3 to 9 cm upward and downward from the inoculation site. Pycnidia were present on collapsed stem tissue, and leaves became chlorotic. Damage was limited to 2mm from the mock-inoculation site, with no discoloration in the control plants. Symptomatic tissues were plated and compared to the original isolates. Alpha conidia were ellipsoidal with the base end rounded. To our knowledge, this is the first report of Diaporthe longicolla causing disease on mungbean within the U.S. and worldwide. The presence of this disease in two locations suggests the potential for Diaporthe longicolla to be a serious disease of mungbean in the future.

Risk assessment for femoral head collapse in osteonecrosis utilizing MRI-derived large lesion ratio: a retrospective cohort study

PurposeThis study aimed to developed a novel and practical method to quantify the involvement of lesion in osteonecrosis of the femoral head (ONFH). We hypothesized that the new metric large lesion ratio (LLR) had promising prognostic value.MethodsA total of 131 hips with non-traumatic ONFH were included in this retrospective study. Patient aged 18–60 with MRI-confirmed diagnosis, and a minimum of 2-year follow-up or radiographic collapse progression during follow-up were included. Patients with prior hip surgery, incomplete data or advanced ONFH at baseline (femoral head collapse > 2 mm or osteoarthritis) were excluded. Involvement of necrotic lesion was evaluated by calculating LLR. The differences of LLR between collapse progression and non-progression groups were investigated, and the differences among different scanning parameters groups were also examined. Prognostic value of LLR was examined by multivariate regression analysis. Receiver operating characteristic curves (ROC) were constructed and areas under the curve (AUC) were compared.ResultsThe median of LLR was 66.67% in the collapse progression group, which was significantly higher compared with 25.00% in the non-progression group (P < 0.001). Subgroups analysis showed that LLR were significantly higher in the collapse progression group of Japanese Investigation Committee type C1 (P < 0.001)and C2 (P = 0.002). Multivariate regression showed that LLR were independently correlated with collapse progression (OR, 1.46 [95% CI, 1.24–1.78]; P < 0.001). ROC analysis showed that the AUC for LLR was 0.84, outperforming the 0.74 AUC OF the JIC classification.ConclusionLLR could served as a efficient tool to assess the risk of collapse progression and guide the selection of treatment strategy.

Severe Antiphospholipid Syndrome in a Child with Chicken Pox: Clinical Case

Background. Antiphospholipid syndrome develops at autoimmune diseases, pregnancy, or genetic predisposition, and it is characterized by high risk of thrombosis in arterial, venous, and capillary vessels. This syndrome in children is rare, and necrotic skin lesions occur in the most extreme cases. Clinical case description. A child, 1 year old, with chicken pox has developed hemorrhagic rash on his back, swelling of the left shoulder and forearm on the 4th day of the disease. Medical treatment included acyclovir, ceftriaxone, sodium heparin. Aggravation of hemorrhagic rash over the body, spread of edema from the left shoulder to upper arm and neck, formation of flaccid bubble and necrosis locus (75 cm) on the outer upper third of the left shoulder were noted on the 5th-6th day. Increase in the levels of antibodies to 2-glycoprotein I (IgG — 48.9 U/ml, IgM — &gt; 100 U/ml) and cardiolipin (IgM — 73.0 U/ml) was revealed. The diagnosis “antiphospholipid syndrome” was established. All necrotized tissues were removed on the 11th day, and the wound defect (86 cm) was subsequently covered by mesh skin grafts. The levels of antibodies (IgG + IgM) to 2-glycoprotein I remained high, 45.2 U/mL, 12 weeks after the onset of the disease. Thus, it has confirmed the development of antiphospholipid syndrome. The child continued treatment with warfarin (3.75 mg/day) with INR control. Conclusion. Hemorrhagic skin lesions in children with infectious diseases requires laboratory diagnostics of antiphospholipid syndrome, early prescription of anticoagulant and antiplatelet therapy, and surgical support.

High-yield, plant-based production of an antimicrobial peptide with potent activity in a mouse model.

Plants offer a promising chassis for the large-scale, cost-effective production of diverse therapeutics, including antimicrobial peptides (AMPs). However, key advances will reduce production costs, including simplifying the downstream processing and purification steps. Here, using Nicotiana benthamiana plants, we present an improved modular design that enables AMPs to be secreted via the endomembrane system and sequestered in an extracellular compartment, the apoplast. Additionally, we translationally fused an AMP to a mutated small ubiquitin-like modifier sequence, thereby enhancing peptide yield and solubilizing the peptide with minimal aggregation and reduced occurrence of necrotic lesions in the plant. This strategy resulted in substantial peptide accumulation, reaching around 2.9 mg AMP per 20 g fresh weight of leaf tissue. Furthermore, the purified AMP demonstrated low collateral toxicity in primary human skin cells, killed pathogenic bacteria by permeabilizing the membrane and exhibited anti-infective efficacy in a preclinical mouse (Mus musculus) model system, reducing bacterial loads by up to three orders of magnitude. A base-case techno-economic analysis demonstrated the economic advantages and scalability of our plant-based platform. We envision that our work can establish plants as efficient bioreactors for producing preclinical-grade AMPs at a commercial scale, with the potential for clinical applications.

First Report of Chili Anthracnose Caused by Colletotrichum sojae in South Korea.

Chili (Capsicum annuum L.) is an economically important crop worldwide, valued for its culinary uses. In South Korea, anthracnose caused by Colletotrichum spp. including C. truncatum, C. gloeosporioides, C. coccodes, C. acutatum, and C. scovillei incurs on substantial economic loss (Kim et al. 2008; Oo and Oh 2020). In August 2022, somewhat different types of symptoms that was not typical on chilli fruits were observed in a field in Yereonggwang (GPS: 35.2579° N, 126.4742° E), South Korea. The disease symptoms appeared as sunken, necrotic lesions with dense black spore masses forming in concentric rings. The estimated disease incidence the 0.2 ha field showing up to 1% of fruits affected. To isolate the pathogen, six symptomatic chilli fruits were collected. Small pieces (5 mm²) were cut from the margins of the lesions, surface-sterilized in 70% ethanol for 30 sec, followed by 1% sodium hypochlorite for 1 minute, and then rinsed three times in sterile distilled water. The tissue pieces were placed on potato dextrose agar (PDA) plates and incubated at 25°C in the dark. After 3 to 5 days, emerging fungal colonies were sub-cultured to obtain pure isolates. A total of five isolates were obtained and initially identified as Colletotrichum spp. based on morphological characteristics. Seven-day old colonies were initially white, turning light orange with age on PDA. Setae (observed on lesion) were dark brown, verruculose and septate. Conidia were cylindrical, hyaline, and measured 14.8 to 19.9 × 4.2 to 6.5 µm (mean 16.7 × 5.6 μm, n = 70) in size; appressoria were brown to dark brown and irregularly shaped. These morphological characteristics of the isolates agree with those reported for the morphology of C. sojae by Damm et al. (2019). To confirm the identity of the isolates, DNA was extracted and specific gene regions were amplified and sequenced using the following primer sets: ITS (ITS1 and ITS4), GAPDH (GDF1 and GDR1), ACT (ACT-512F and ACT-783R), TUB (T1 and Bt2b), HIS3 (CYLH3F and CYLH3R), and CHS-1 (CHS-79F and CHS-345R). The resulting sequences were deposited in the NCBI GenBank with accession numbers (LC830742 to LC830766). Maximum likelihood phylogenetic analysis using combine sequences of ITS, GAPDH, ACT, TUB, HIS3 and CHS-1 in MEGA X confirmed the isolates as C. sojae, marking the first report of this pathogen on chilli in South Korea, previously known to infect soybean. Pathogenicity tests were conducted on wound and nonwounded healthy and mature-green chili fruits (cv. Bicksita) to confirm the pathogenicity of the isolated C. sojae. The fruits were surface-sterilized using 70% ethanol and then rinsed with sterile distilled water. The fruits were wounded using a sterile needle to facilitate infection. A conidial suspension (1x106 conidia/mL) was prepared from 7-day-old PDA cultures. Each fruit was inoculated by placing a 10 µL drop of the conidial suspension onto the wounded and nonwounded sites (4 to 5) of the wound and unwound fruits, respectively. Control fruits were inoculated with sterile water. A total of 40 fruits per treatment were used and the experiment repeated twice. The fruits were placed in plastic box lined with moist paper towels to maintain high humidity and incubated at 25°C. Anthracnose symptoms developed on the inoculated fruits within 7 days, while control and unwounded fruits remained symptom-free. Colletotrichum sojae was successfully reisolated from the symptomatic fruits, fulfilling Koch's postulates and confirming its role as the causal agent of the disease. Colletotrichum sojae is known to infect Fabaceae species worldwide such as Glycine max, Medicago sativa, Phaseolus vulgaris, Atractylodes ovata and Vigna unguiculata (Damm et al. 2019; Talhinhas and Baroncelli 2021), Atractylodes ovata in South Korea (Hassan et al. 2021) and chili pepper in China (Zhanget al. 2023). The first report of C. sojae causing chili anthracnose in South Korea represents a new challenge for chili growers. Integrated disease management strategies need to be developed and implemented to mitigate its impact.

LAMTOR1/mTORC1 promotes CD276 to induce immunosuppression via PI3K/Akt/MMP signaling pathway in Clostridium perfringens–induced necrotic enteritis of laying hens

Clostridium perfringens (C. perfringens) causes avian necrotic enteritis, leading to huge economic losses to the poultry industry. This pathogen induces host immunosuppression; however, the molecular mechanism is still unclear. Thus, we established a laying hen infection model to explore this mechanism. We randomly divided 20 one-old-day laying hens into the control and infection groups. The infection group was infected intragastrically with 1 × 109 colony-forming units of C. perfringens in 1 mL of sterile phosphate-buffered saline (PBS) once a day from d 17 to 20; the control group received the same volume of PBS without the bacterium. Twenty-four hours after the last challenge, we sacrificed the laying hens and collected the jejunum for analysis. The infection group presented alterations in blood biochemical parameters and necrotic lesion scores as well as damage to the jejunum. Proteomics revealed 427 upregulated and 291 downregulated proteins in the infection group. In the infection group, CD3, CD4, and CD8 messenger RNA expression (mRNA) expression was decreased; LAMTOR1 and mTORC1 mRNA expression was increased; CD276 protein expression was enhanced; and the PI3K/Akt/MMP pathway was activated in jejunum of laying hens. This is the first study to report CD276 expression in the jejunum related to immunosuppression in a laying hen model of necrotic enteritis. It provides some new key targets to potentially control avian necrotic enteritis.

Risk Factors for Femoral Head Collapse in Osteonecrosis

BackgroundOsteonecrosis of the femoral head (ONFH) affects at least 20,000 patients annually in the United States; however, the pathophysiology of disease progression is poorly understood. The purpose of this study was to determine the relative importance of three distinct elements and their relationship to the collapse of the femoral head as follows: (1) identifiable risk factors, (2) femoral head anatomy, and (3) the extent of the necrotic lesion. MethodsA single-center retrospective cohort study was performed on patients ≥ 18 year old who presented with ONFH. Ficat classification and femoral head anatomic parameters were measured on radiographs. Osteonecrotic lesion size was measured on magnetic resonance imaging using four validated methods. Multivariable regression analyses were performed to identify predictors of femoral head collapse. ResultsThere were 105 patients and 137 hips included in the final cohort, of which 50 (36.5%) had collapse of the femoral head. Multivariable analyses demonstrated that medical risk factors (adjusted odds ratio (aOR): 1.15), alcohol exposure (aOR: 1.23), and increased alpha angle (aOR: 4.51) were predictive of femoral head collapse. Increased femoral head offset (aOR: 0.54) was protective against collapse. An increased size of the osteonecrotic lesion was significantly predictive of collapse with all four measurement methods evaluated: three-dimensional volumetric (aOR: 3.73), modified Kerboul (aOR: 2.92), index of necrotic extent (aOR: 1.91), and modified index of necrotic extent (aOR: 2.05). ConclusionsIn an analysis of patients who had ONFH, we identified risk factors such as alcohol exposure, high alpha angle, increased lesion size, and decreased femoral offset as increasing the risk of femoral head collapse. Given the challenges of studying this patient population, large prospective studies of patients who have ONFH should seek to identify whether these factors are reliable indicators of femoral head collapse. Level of EvidenceIII.

First Report of Colletotrichum queenslandicum Causing Leaf Anthracnose on Jatropha curcas L in China.

Jatropha curcas L. (Euphorbiaceae), is a valuable multi-purpose crop, previously used for disease treatment and environmental restoration, recently is attention to use jatropha oil for produce biodiese. In June 2023, the leaf disease of approximately 60% of J. curcas was observed in Mazhang, Zhanjiang, Guangdong Province (E110°27'26.8'' N22°6'14.7''). Diseased leaves showed typical anthracnose symptoms of chlorotic regions with brownish sunken necrotic lesions (Figure 1). Sections from the junction of disease were surface disinfected in 75% ethanol and 3% hydrogen peroxide solution for 1 minute each. Four small pieces of infected tissue were removed from the lesion and placed on potato dextrose agar (PDA), and incubated at 25 to 28℃ in the dark. Hyphal tips from the inoculated tissues were subcultured on PDA and two isolates were purified by single spore method. The colonies on PDA were 7.8 cm diam after 10 d at 25 to 28 ℃, covered with dense, cottony, grayish-white aerial mycelium and small dark-based acervuli with orange ooze and dark brown straight setae. Conidia were hyaline, smooth-walled, aseptate, the apex and base rounded, slightly constricted near centre, 12.9 - 13.8 × 3.9 - 4.6 um (av.13.6 × 4.3 μm, n = 50). Appressoria were variable in shape, mostly simple, subglobose or irregular lobes, 5.8-9.6 × 5.7-11.2 um (Figure 2). Perithecia were not observed. These characteristics were consistent with Colletotrichum sp. (Weir, B. S., et al. 2012). Sequences of isolates ACCC 35630 and ACCC 35631 stored in Agricultural Culture Collection of China including internal transcribed spacer (ITS), actin (ACT), beta-tubulin (tub2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and chitin synthase (chs). were amplified (Weir et al, 2012), sequenced and submitted to GenBank (ITS: PP474979 and PP474984; ACT: PP505487 and PP505488; TUB: PP505493 and PP505494; GAPDH: PP505491 and PP505492; CHS: PP505489 and PP505490). The amplicon sizes of ITS, ACT, TUB, GAPDH, and CHS were 550, 652, 500, 264, and 301 bp, respectively. Phylogenetic analyses showed that Isolates 35630 & 35631 were clustered closely association with RHCOL1 and RHCOL3. Phylogenetic analysis with MEGA 7 using the combined ITS-ACT-CHS-GAPDH-TUB2 sequences showed that the two isolates clustered with C. queenslandicum (Figure 3). To test the pathogenicity, ten healthy leaves on plants in the field were wiped with 75% alcohol and sterile water, punctured with a sterile needle and inoculated by adding 10 uL of spore suspension (1 × 105 conidia/ml) to the wounded sites. And two other leaves were added sterile water as controls. Symptoms of anthracnose were observed on leaves similar to the disease described above after 7 days of inoculation, whereas the leaves from the controls remained asymptomatic. C. queenslandicum was reisolated from the inoculated leaves. C. queenslandicum has been reported as a pathogen causing leaf and fruit anthracnose on papaya, coffee, rambutan, avocado and Persian lime etc. in tropical and temperate regions (Kunta, M., et al. 2018), and this is the first report on J. curcas in China as well as worldwide. This disease may have a significant negative impact on J. curcas cultivation.

Characterization of mycotoxins produced by two Fusarium species responsible for postharvest rot of banana fruit

AbstractIn an open-air market in southern Italy, we noticed ‘Lady finger’ banana fruit imported from Costa Rica showing a severe rot, whose symptoms consisted of necrotic peel lesions with variable shape and size. Fusarium sacchari and F. proliferatum were consistently isolated from symptomatic fruit. In pathogenicity tests on ‘Lady finger’ banana fruit, F. proliferatum was more virulent than F. sacchari. Quantitative Time-of-Flight Mass Spectrometric analysis of secondary metabolites produced by isolates of these two Fusarium species on three different matrices (banana peel, barley and maize kernels) identified 11 mycotoxins. Seven of them (Fusaproliferin, Fumonisins A1, Fumonisins A2 and Fumonisins B1, Hydrolysed Fumonisin B1, Fusarin C and Moniliformin) were detected in matrices contaminated by F. proliferatum isolates. Fumonisin A1 was the prevalent mycotoxin in both maize kernels and banana peel, while Fumonisin A2 prevailed in barley kernels. Similarly, seven mycotoxins (the cyclic hexadepsipeptides Enniatins B2, B3 and B4, Fumonisins A1 and B2, Hydrolysed Fumonisin B1 and Fusarin C) were detected in matrices contaminated by F. sacchari isolates, but they were only in part the same as those produced by F. proliferatum isolates. Fusarin C prevailed in all three matrices colonized by F. sacchari. Fumonisin A1 was detected exclusively in maize kernels while Enniatins B3 and B4, Fumonisin B2 and Hydrolysed Fumonisin B1 were detected exclusively in barley kernels. Overall, F. proliferatum produced a higher amount of mycotoxins than F. sacchari. Moreover, in banana peel both species produced a lower number and amount of mycotoxins than in the other two matrices.

Violaceous nodules of the extensor joints - a clinicopathological challenge.

Antineutrophilic cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is a condition characterized by vessel inflammation and may have a variety of etiologies. Among these, cocaine and its common adulterant, levamisole, have been described to contribute to the development of AAV with distinct cutaneous manifestations. Classically, these manifestations involve purpuric or necrotic lesions involving the ears, nose, and extremities. However, we present a case of cocaine-induced AAV presenting with violaceous nodules on the dorsal hands in order to demonstrate that this condition may not always present with retiform purpura and skin necrosis.